| 禽白血病病毒衣壳蛋白p27基因的原核表达及多克隆抗体的制备 |
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| 引用本文： | 李德龙,刘在斯,贠炳岭,吴关,高玉龙,秦立廷,祁小乐,王永强,高宏雷,刘思当,王笑梅.禽白血病病毒衣壳蛋白p27基因的原核表达及多克隆抗体的制备[J].中国家禽,2012(9):13-16. |
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| 作者姓名： | 李德龙 刘在斯 贠炳岭 吴关 高玉龙 秦立廷 祁小乐 王永强 高宏雷 刘思当 王笑梅 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所;山东农业大学 |
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| 基金项目： | 现代农业肉鸡产业技术体系建设(nycytx-42-G3-01);国家自然科学基金(31072146);哈尔滨市科技攻关计划项目(2010AA6AN034) |
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| 摘 要： | 从J亚群禽白血病病毒(ALV-J)HB09JY03株感染的DF-1细胞的冻融裂解液中提取DNA,通过PCR方法扩增出约720bp的gag-p27基因片段,克隆至pMD18-T载体。鉴定正确后,将该片段克隆至pET-30a(+)原核表达载体,经IPTG诱导表达。表达产物经His Trap TMHP纯化后,测蛋白浓度达到9mg/mL。用兔抗自然p27蛋白阳性血清进行Western blot检测,表明重组p27蛋白有抗原反应活性。免疫新西兰大白兔后,产生的抗体与禽白血病全病毒抗原可以发生特异性反应。所制备的重组p27蛋白和多克隆抗体将在ALV的抗原分析、血清学诊断等方面有着重要的应用价值。
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| 关 键 词： | 禽白血病 p27基因 原核表达 多克隆抗体 |
Expression of Capsid Protein p27 Gene of Avian Leucosis Virus and Preparation of Multiclonal Antibodies Against Recombinant p27 Protein |
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| LI Delong,LIU Zaisi,YUN Bingling,WU Guan,GAO Yulong,QIN Liting,QI Xiaole,WANG Yongqiang,GAO Honglei,LIU Sidang,Wang Xiaomei.Expression of Capsid Protein p27 Gene of Avian Leucosis Virus and Preparation of Multiclonal Antibodies Against Recombinant p27 Protein[J].China Poultry,2012(9):13-16. |
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| Authors: | LI Delong LIU Zaisi YUN Bingling WU Guan GAO Yulong QIN Liting QI Xiaole WANG Yongqiang GAO Honglei LIU Sidang Wang Xiaomei |
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| Institution: | 1(1.Harbin Veterinary Research Institute of CAAS,Harbin,Heilongjiang 150001;2.Shandong Agricultural University,Shandong Tai’an,Shandong 271000) |
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| Abstract: | DNA was extracted from DF-1 cell lysate infected with subgroup J of avian leucosis virus(ALV) and the gag-p27 gene was amplified by PCR.Then the gene was cloned into plasmid pMD18-T and sequenced.After identified exactly,the whole coding fragment of p27 gene was cloned into prokaryotic expression vector pET-30a(+) and induced with IPTG.After purification with His TrapTM HP,the density of p27 recombinant protein was up to 9 mg/mL.The immune reactivity of recombinant p27 protein was identified with positive antiserum against nature p27 protein specifically by Western-blot.Multiclonal antibodies against recombinant p27 protein was obtained by subcutaneous injection of rabbit,which is specific to ALV antigen and it would be very useful as a tool for antigenic analysis and serological diagnosis of ALV. |
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| Keywords: | avian leucosis p27 gene prokaryotic expression multiclonal antibodies |
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